卡氏肺孢子主要表面糖蛋白UCS基因siRNA表达载体的构建和鉴定  

Construction and identification of plasmid vector of short interfering RNA on pneumocystis carinii major surface glycoprotein gene's upstream conserved sequence

作　　者：谭燕[1] 姚云清[1] 王莉妮[1] 唐滢浍[1] 李文桂[1] 马良 陈雅棠[1] 

机构地区：[1]重庆医科大学附属第一医院感染科,重庆400016 [2]美国路易斯安那州新奥尔良大学医学院,路易斯安那州70112

出　　处：《重庆医科大学学报》2008年第3期264-266,共3页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(30471512);重庆市医学科技计划项目(04-2-100)。

摘　　要：目的:构建并鉴定艾滋病主要机会性感染病原体-卡氏肺孢子(Pneumocystis carinii,PC)主要表面糖蛋白(Major surface glycoprotein,MSG)表达调控基因上游保守序列(Upstream conserved sequence,UCS)的短干扰性RNA(Short interfering RNA,siRNA)表达载体。方法:设计并人工合成针对PC MSG-UCS基因的短发夹状(Short hairpin RNA,shRNA)序列并定向克隆到siRNA表达载体pTZU6+1上,采用双酶切与测序法进行鉴定。结果:酶切和测序鉴定得到的产物与预期目的基因一致。结论:成功构建和鉴定PC MSG-UCS基因的siRNA表达载体。Objective： To construct and identify a plasmid vector of short interfering RNA（siRNA） on pneumocystis carinii （PC） major surface glycoprotein （ MSG ） gene＇ s upstream conserved sequence （ UCS ）. Methods ： Short hairpin RNA （ shRNA ） oligonucleotide targeting PC MSG UCS gene which was chemically synthesized and inserted into RNAi-Ready plasmid vector pTZU6＋1 after annealing. The recombinant plasmid, PC-UCS, transformed into E. coil. TOP10 and amplified,was digested by restriction endonucleases HindlII and EcoRI and identified by gel electrophoresis and DNA sequencing. Results：Gel electrophoresis and DNA sequencing showed that the recombinant plasmid containing the correct and full shRNA oligonucleotide. Conclusion：The siRNA plasmid,pPC-UCS, was constructed successfully.

关 键 词：PC MSG-UCS SIRNA 表达载体 

分 类 号：R730.23[医药卫生—肿瘤]